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Abstract
Extravillous trophoblast cells (EVTs) secreted by the uterine cavity may help overcome limitations associated with prenatal testing currently in use. EVTs are isolated using a routine safe liquid-based Pap test (called ThinPrep); however, the ThinPrep solution contains alcohol that hinders the isolation of intact EVTs. We compared the trophoblastic cell isolation efficiency of two different methods of fixation: Thinprep (pre-fixation method) and formalin (post-fixation method). We analyzed EVTs from 20 pregnant women (5–20 weeks of gestation) who underwent invasive prenatal testing. The percentages of placental β-human chorionic gonadotropin (β-hCG)-expressing cells were calculated. The presence of XY chromosomes were used to confirm pure trophoblast cells by fluorescence in situ hybridization. The β-hCG-positive cells obtained from pre- and post-fixation were 66.4 ± 13.3% and 83.2 ± 8.1% (p = 0.003), respectively, and fluorescence-positive cells were 11.1 ± 2.1% and 23.8 ± 4.8%, respectively (p = 0.001). Post-fixation was found to be more efficient in isolating non-trophoblast cells than pre-fixation. For the successful clinical application of trophoblast retrieval and isolation from the cervix in prenatal genetic testing, each step should be optimized for consistent and reliable results.
Highlights
The current average age of mothers at childbirth is over 30 years in developed countries due to women pursuing higher education and developing their careers, women marrying at an older age, and advancements in assisted reproductive technology [1]
fluorescence in situ hybridization (FISH) revealed positive rates of 11.1 ± 2.1% (8.6%–13.5%) and 23.8 ± 4.8% (19.4%–29.4%) after pre- and post-fixation, respectively (p = 0.001) (Table 3)
The reproducibility of TRIC remains controversial due to its 75%–100% success rates in confirming fetal tissues using β-human chorionic gonadotropin (β-hCG) [14]. This limits the clinical application of TRIC as a prenatal diagnostic tool
Summary
The current average age of mothers at childbirth is over 30 years in developed countries due to women pursuing higher education and developing their careers, women marrying at an older age, and advancements in assisted reproductive technology [1]. The birth rate for women aged 35–44 years steadily increased from 5.2% to 15.5% in 2014 [2]. Emphasis should be put on the importance of prenatal genetic testing because the risk of fetal chromosomal abnormalities increases in childbearing women of advanced maternal age of >35 years [3]. Confirmatory prenatal genetic tests, such as chorionic villus sampling (CVS) and amniocentesis, are invasive procedures. These tests are associated with a procedure-related miscarriage rate of 0.1–2%. That increases maternal anxiety [4,5]. Patients and healthcare providers require noninvasive and Diagnostics 2020, 10, 300; doi:10.3390/diagnostics10050300 www.mdpi.com/journal/diagnostics
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